Copello J, Simon B, Segal Y, Wehner F, Ramanujam V M, Alcock N, Reuss L
Department of Physiology, University of Texas Medical Branch, Galveston 77550.
Biophys J. 1991 Oct;60(4):931-41. doi: 10.1016/S0006-3495(91)82127-0.
Glasses used to fabricate patch pipettes may release components which affect ion channels (Cota, G., and C.M. Armstrong. 1988. Biophys. J. 53:107-109; Furman, R.E., and J.C. Tanaka. 1988. Biophys. J. 53:287-292; Rojas, L., and C. Zuazaga. 1988. Neurosci. Lett. 88:39-44). The gating properties of maxi K+ channels from Necturus gallbladder epithelium depend on whether borosilicate glass (BG) or blue tip hematocrit glass (SG) is used to construct the patch pipettes. The data are consistent with solubilization from SG of a component which exerts voltage-dependent, cytosolic-side specific block, closely resembling "slow block" by Ba2+ ions. Ringer's solution preincubated with SG, but not with BG, blocked inside-out maxi K+ channels when used as bathing solution. Mass spectrometry revealed that Ba2+ is released by the glass from fast and slow-release compartments (SG contains 3% wt/wt BaO), and is the only ion found in the solution at concentrations consistent with the observed channel block. Additionally, SG released O2-, Na+, Ca2+, and Mg2+, all to micromolar concentrations. These elements do not interfere with maxi K+ channels but they could in principle alter the properties of other ion channels. Thus, screening for channel-modifying substances released by the glass may be necessary for the adequate interpretation of patch-clamp results.
用于制作膜片钳微电极的玻璃可能会释放影响离子通道的成分(科塔,G.,和 C.M. 阿姆斯特朗。1988 年。《生物物理学杂志》53:107 - 109;弗曼,R.E.,和 J.C. 田中。1988 年。《生物物理学杂志》53:287 - 292;罗哈斯,L.,和 C. 苏阿萨加。1988 年。《神经科学快报》88:39 - 44)。美西螈胆囊上皮细胞大电导钾通道的门控特性取决于用于构建膜片钳微电极的是硼硅酸盐玻璃(BG)还是蓝尖血细胞比容玻璃(SG)。数据表明,SG 可溶解一种成分,该成分能产生电压依赖性的、胞质侧特异性阻断,与 Ba²⁺离子引起的“慢阻断”极为相似。当用作浴液时,用 SG 预孵育但不用 BG 预孵育的林格氏液可阻断内向外模式的大电导钾通道。质谱分析显示,玻璃从快速和缓慢释放区室释放出 Ba²⁺(SG 含有 3%重量/重量的 BaO),并且是溶液中发现的唯一浓度与观察到的通道阻断一致的离子。此外,SG 还释放出 O²⁻、Na⁺、Ca²⁺和 Mg²⁺,浓度均为微摩尔级。这些元素不会干扰大电导钾通道,但原则上可能会改变其他离子通道的特性。因此,为了充分解释膜片钳实验结果,可能有必要筛选玻璃释放的通道修饰物质。